Anti-FOXO3A antibody (ab23683)

Overview

  • Product nameAnti-FOXO3A antibody
    See all FOXO3A primary antibodies
  • Description
    Rabbit polyclonal to FOXO3A
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Human FOXO3A.

    (Peptide available as ab24031.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: Jurkat (Human T cell lymphoblast-like cell line) This antibody gave a positive signal in the following tissue lysates: Mouse Heart Mouse Skeletal Muscle Rat Heart This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab23683 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionTranscriptional activator which triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. Recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3'.
  • Tissue specificityUbiquitous.
  • Involvement in diseaseNote=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
  • Sequence similaritiesContains 1 fork-head DNA-binding domain.
  • Post-translational
    modifications
    In the presence of survival factors such as IGF-1, phosphorylated on Thr-32 and Ser-253 by AKT1/PKB. This phosphorylated form then interacts with 14-3-3 proteins and is retained in the cytoplasm. Survival factor withdrawal induces dephosphorylation and promotes translocation to the nucleus where the dephosphorylated protein induces transcription of target genes and triggers apoptosis. Although AKT1/PKB doesn't appear to phosphorylate Ser-315 directly, it may activate other kinases that trigger phosphorylation at this residue. Phosphorylated by STK4 on Ser-209 upon oxidative stress, which leads to dissociation from YWHAB/14-3-3-beta and nuclear translocation. Phosphorylated by PIM1.
  • Cellular localizationCytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors.
  • Information by UniProt
  • Database links
  • Alternative names
    • AF6q21 antibody
    • AF6q21 protein antibody
    • DKFZp781A0677 antibody
    • FKHR2 antibody
    • FKHRL 1 antibody
    • FKHRL1 antibody
    • FKHRL1P2 antibody
    • Forkhead (Drosophila) homolog (rhabdomyosarcoma) like 1 antibody
    • Forkhead box O3 antibody
    • Forkhead box O3A antibody
    • Forkhead box protein O3 antibody
    • Forkhead box protein O3A antibody
    • Forkhead Drosophila homolog of in rhabdomyosarcoma like 1 antibody
    • Forkhead homolog (rhabdomyosarcoma) like 1 antibody
    • Forkhead in rhabdomyosarcoma like 1 antibody
    • Forkhead in rhabdomyosarcoma-like 1 antibody
    • FOX O3A antibody
    • FOXO2 antibody
    • foxo3 antibody
    • FOXO3_HUMAN antibody
    • FOXO3A antibody
    • MGC12739 antibody
    • MGC31925 antibody
    see all

Anti-FOXO3A antibody images

  • All lanes : Anti-FOXO3A antibody (ab23683) at 1 µg/ml

    Lane 1 : Jurkat whole cell lysate (ab7899)
    Lane 2 : Jurkat whole cell lysate (ab7899) with Human FOXO3A peptide (ab24031) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.


    Predicted band size : 71 kDa
    Observed band size : 90 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 50 kDa (possible cleavage fragment,cross reactivity).
  • All lanes : Anti-FOXO3A antibody (ab23683) at 1 µg/ml

    Lane 1 : Heart (Mouse) Tissue Lysate - normal tissue
    Lane 2 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 3 : Heart (Rat) Tissue Lysate - normal tissue

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size : 71 kDa
    Observed band size : 90 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 16 kDa,27 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab23683 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23683, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% HepG2 fixed (10 min) HepG2 cells at 5µg/ml.
  • IHC image of FOXO3A staining in Human normal lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab23683, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-FOXO3A antibody (ab23683)

ab23683 has not yet been referenced specifically in any publications.

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Thank you for your reply. I do not believe there will be any problems with the antibody being in a different tube but please let me know if the customer has issues with the antibody being packaged different. I will keep you posted when I hear anything.

I am unsure as to why ab23683 was shipped in a 1ml vial instead of a 0.5ml vial. I am in the process of trying to get an answer for you. I was wandering though if you feel the quality of the product is being affected by it being shipped in a different ...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon)
Specification Colon
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Dec 16 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Jurkat)
Total protein in input 400 µg
Specification Jurkat
Immuno-precipitation step Protein A
Username

Mr. Antibody Solutions

Verified customer

Submitted Apr 28 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"